+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 4cg5 | ||||||
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タイトル | Cryo-EM of the Sec61-complex bound to the 80S ribosome translating a secretory substrate | ||||||
要素 |
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キーワード | PROTEIN TRANSPORT / RIBOSOME / CO-TRANSLATIONAL PROTEIN TRANSLOCATION | ||||||
機能・相同性 | 機能・相同性情報 Insertion of tail-anchored proteins into the endoplasmic reticulum membrane / membrane docking / endoplasmic reticulum Sec complex / pronephric nephron development / cotranslational protein targeting to membrane / Ssh1 translocon complex / Sec61 translocon complex / protein targeting to ER / protein-transporting ATPase activity / protein insertion into ER membrane ...Insertion of tail-anchored proteins into the endoplasmic reticulum membrane / membrane docking / endoplasmic reticulum Sec complex / pronephric nephron development / cotranslational protein targeting to membrane / Ssh1 translocon complex / Sec61 translocon complex / protein targeting to ER / protein-transporting ATPase activity / protein insertion into ER membrane / SRP-dependent cotranslational protein targeting to membrane, translocation / signal sequence binding / post-translational protein targeting to membrane, translocation / protein transmembrane transporter activity / guanyl-nucleotide exchange factor activity / phospholipid binding / ribosome binding / endoplasmic reticulum membrane / endoplasmic reticulum 類似検索 - 分子機能 | ||||||
生物種 | CANIS LUPUS FAMILIARIS (イヌ) | ||||||
手法 | 電子顕微鏡法 / 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 7.4 Å | ||||||
データ登録者 | Gogala, M. / Becker, T. / Beatrix, B. / Barrio-Garcia, C. / Berninghausen, O. / Beckmann, R. | ||||||
引用 | ジャーナル: Nature / 年: 2014 タイトル: Structures of the Sec61 complex engaged in nascent peptide translocation or membrane insertion. 著者: Marko Gogala / Thomas Becker / Birgitta Beatrix / Jean-Paul Armache / Clara Barrio-Garcia / Otto Berninghausen / Roland Beckmann / 要旨: The biogenesis of secretory as well as transmembrane proteins requires the activity of the universally conserved protein-conducting channel (PCC), the Sec61 complex (SecY complex in bacteria). In ...The biogenesis of secretory as well as transmembrane proteins requires the activity of the universally conserved protein-conducting channel (PCC), the Sec61 complex (SecY complex in bacteria). In eukaryotic cells the PCC is located in the membrane of the endoplasmic reticulum where it can bind to translating ribosomes for co-translational protein transport. The Sec complex consists of three subunits (Sec61α, β and γ) and provides an aqueous environment for the translocation of hydrophilic peptides as well as a lateral opening in the Sec61α subunit that has been proposed to act as a gate for the membrane partitioning of hydrophobic domains. A plug helix and a so-called pore ring are believed to seal the PCC against ion flow and are proposed to rearrange for accommodation of translocating peptides. Several crystal and cryo-electron microscopy structures revealed different conformations of closed and partially open Sec61 and SecY complexes. However, in none of these samples has the translocation state been unambiguously defined biochemically. Here we present cryo-electron microscopy structures of ribosome-bound Sec61 complexes engaged in translocation or membrane insertion of nascent peptides. Our data show that a hydrophilic peptide can translocate through the Sec complex with an essentially closed lateral gate and an only slightly rearranged central channel. Membrane insertion of a hydrophobic domain seems to occur with the Sec complex opening the proposed lateral gate while rearranging the plug to maintain an ion permeability barrier. Taken together, we provide a structural model for the basic activities of the Sec61 complex as a protein-conducting channel. | ||||||
履歴 |
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-構造の表示
ムービー |
ムービービューア |
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構造ビューア | 分子: MolmilJmol/JSmol |
-ダウンロードとリンク
-ダウンロード
PDBx/mmCIF形式 | 4cg5.cif.gz | 103.5 KB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb4cg5.ent.gz | 76.7 KB | 表示 | PDB形式 |
PDBx/mmJSON形式 | 4cg5.json.gz | ツリー表示 | PDBx/mmJSON形式 | |
その他 | その他のダウンロード |
-検証レポート
文書・要旨 | 4cg5_validation.pdf.gz | 806.2 KB | 表示 | wwPDB検証レポート |
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文書・詳細版 | 4cg5_full_validation.pdf.gz | 815.8 KB | 表示 | |
XML形式データ | 4cg5_validation.xml.gz | 22.6 KB | 表示 | |
CIF形式データ | 4cg5_validation.cif.gz | 32.4 KB | 表示 | |
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/cg/4cg5 ftp://data.pdbj.org/pub/pdb/validation_reports/cg/4cg5 | HTTPS FTP |
-関連構造データ
-リンク
-集合体
登録構造単位 |
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-要素
#1: タンパク質 | 分子量: 52279.379 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) CANIS LUPUS FAMILIARIS (イヌ) / 器官: PANCREAS / 参照: UniProt: P38377 |
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#2: タンパク質 | 分子量: 7752.325 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) CANIS LUPUS FAMILIARIS (イヌ) / 器官: PANCREAS / 参照: UniProt: P60058 |
#3: タンパク質・ペプチド | 分子量: 3992.791 Da / 分子数: 1 / Fragment: RESIDUES 61-96 / 由来タイプ: 天然 / 由来: (天然) CANIS LUPUS FAMILIARIS (イヌ) / 器官: PANCREAS / 参照: UniProt: P60467 |
-実験情報
-実験
実験 | 手法: 電子顕微鏡法 |
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EM実験 | 試料の集合状態: PARTICLE / 3次元再構成法: 単粒子再構成法 |
-試料調製
構成要素 | 名称: CANIS FAMILIARIS SEC61 BOUND TO A WHEAT GERM 80S-RNC TRANSLATING THE TRANSLOCATING LEPT-POLYPEPTIDE タイプ: RIBOSOME |
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緩衝液 | 名称: 30 MM HEPES/KOH 7.6, 10 MM MG(OAC)2, 180 MM KOAC/HAC PH 7.6, 0.3 % DIGITONIN, 1 MM DTT pH: 7.6 詳細: 30 MM HEPES/KOH 7.6, 10 MM MG(OAC)2, 180 MM KOAC/HAC PH 7.6, 0.3 % DIGITONIN, 1 MM DTT |
試料 | 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES |
試料支持 | 詳細: CARBON |
急速凍結 | 装置: FEI VITROBOT MARK IV / 凍結剤: ETHANE 詳細: VITRIFICATION 1 -- CRYOGEN- ETHANE, HUMIDITY- 95, INSTRUMENT- FEI VITROBOT MARK IV, METHOD- BLOT FOR 3 SECONDS BEFORE PLUNGING, |
-電子顕微鏡撮影
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |
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顕微鏡 | モデル: FEI TITAN KRIOS / 日付: 2011年7月17日 |
電子銃 | 電子線源: FIELD EMISSION GUN / 加速電圧: 200 kV / 照射モード: FLOOD BEAM |
電子レンズ | モード: BRIGHT FIELD / 倍率(補正後): 148721 X / 最大 デフォーカス(公称値): 4000 nm / 最小 デフォーカス(公称値): 1300 nm / Cs: 2.7 mm |
撮影 | 電子線照射量: 25 e/Å2 フィルム・検出器のモデル: TVIPS TEMCAM-F416 (4k x 4k) |
画像スキャン | デジタル画像の数: 9805 |
放射波長 | 相対比: 1 |
-解析
EMソフトウェア |
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CTF補正 | 詳細: ON 3D-VOLUME (SPIDER) | |||||||||||||||||||||
対称性 | 点対称性: C1 (非対称) | |||||||||||||||||||||
3次元再構成 | 解像度: 7.4 Å / 粒子像の数: 53248 / ピクセルサイズ(公称値): 1.2375 Å 詳細: SUBMISSION BASED ON EXPERIMENTAL DATA FROM EMDB EMD-2511. (DEPOSITION ID: 12121). 対称性のタイプ: POINT | |||||||||||||||||||||
原子モデル構築 | プロトコル: FLEXIBLE FIT / 空間: REAL / 詳細: METHOD--FLEXIBLE | |||||||||||||||||||||
原子モデル構築 | PDB-ID: 2WWB Accession code: 2WWB / Source name: PDB / タイプ: experimental model | |||||||||||||||||||||
精密化 | 最高解像度: 7.4 Å | |||||||||||||||||||||
精密化ステップ | サイクル: LAST / 最高解像度: 7.4 Å
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